A Novel 3D In Vitro Platform for Pre-Clinical Investigations in Drug Testing, Gene Therapy, and Immuno-oncology
A Novel 3D In Vitro Platform for Pre-Clinical Investigations in Drug Testing, Gene Therapy, and Immuno-oncology
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DOI：
10.1038/s41598-019-43613-9
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发表时间：
2019-05-09
影响因子：
4.6
通讯作者：
Dominici, Massimo
中科院分区：
文献类型：
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作者：
Candini, Olivia;Grisendi, Giulia;Dominici, Massimo
肿瘤在复杂的细胞间相互作用中发展，辅助细胞从癌症进展的早期阶段开始发挥相关作用。这一事件发生在三维(3D)环境中，到目前为止，由于其复杂性，在体外很难复制。虽然二维培养已经产生了大量数据，但人们逐渐意识到，3D培养策略可以迅速增加对肿瘤发展的理解，并用于抗癌化合物筛选和利用个性化方法预测新药的反应。然而，需要能够在体外3D快速重建癌症组织的简单系统，并且可以用于各种应用。因此，我们开发了一种平板、手持和多功能的3D细胞培养生物反应器，它可以装载肿瘤和/或正常细胞的组合，可以使用各种读数进行监测。这种生物兼容设备支持代表各种癌症的肿瘤细胞系的3D生长，如胰腺癌和乳房腺癌、肉瘤和胶质母细胞瘤。细胞重新填充被两层透气膜完全与外层空间隔开的薄基质，并使用显微镜和光度学对其进行实时监测，即使在运输后也是如此。该设备在3D细胞毒性测试中进行了测试，以研究联合培养中化疗、生物制剂和基于细胞的治疗的抗癌潜力。在靶癌细胞中加入荧光素酶适合于比较研究，也可能涉及平行的活体研究。值得注意的是，该系统受到了挑战，使用从肺癌患者收集的原代肿瘤细胞作为一种创新的预测癌症对检查点抑制剂(如nivolumab)的反应性的功能分析。这种生物反应器在3D培养研究领域具有几个新颖的特征，代表了一种有效的工具，可用于癌症调查、药物筛选和其他毒理学方法。
Tumors develop within complex cell-to-cell interactions, with accessory cells playing a relevant role starting in the early phases of cancer progression. This event occurs in a three-dimensional (3D) environment, which to date, has been difficult to reproduce in vitro due to its complexity. While bi-dimensional cultures have generated substantial data, there is a progressive awareness that 3D culture strategies may rapidly increase the understanding of tumor development and be used in anti-cancer compound screening and for predicting response to new drugs utilizing personalized approaches. However, simple systems capable of rapidly rebuilding cancer tissues ex-vivo in 3D are needed and could be used for a variety of applications. Therefore, we developed a flat, handheld and versatile 3D cell culture bioreactor that can be loaded with tumor and/or normal cells in combination which can be monitored using a variety of read-outs. This biocompatible device sustained 3D growth of tumor cell lines representative of various cancers, such as pancreatic and breast adenocarcinoma, sarcoma, and glioblastoma. The cells repopulated the thin matrix which was completely separated from the outer space by two gas-permeable membranes and was monitored in real-time using both microscopy and luminometry, even after transportation. The device was tested in 3D cytotoxicity assays to investigate the anti-cancer potential of chemotherapy, biologic agents, and cell-based therapy in co-cultures. The addition of luciferase in target cancer cells is suitable for comparative studies that may also involve parallel in vivo investigations. Notably, the system was challenged using primary tumor cells harvested from lung cancer patients as an innovative predictive functional assay for cancer responsiveness to checkpoint inhibitors, such as nivolumab. This bioreactor has several novel features in the 3D-culture field of research, representing a valid tool useful for cancer investigations, drug screenings, and other toxicology approaches.